Assay of aspartylglycosylaminase by high-performance liquid chromatography.
An aspartylglycosylaminase assay based on high-performance liquid chromatographic analysis of the substrate aspartylglucosamine and product aspartate is described. Aspartylglucosamine and aspartate are derivatized with phenylisothiocyanate and resolved by reverse-phase chromatography. The detection limit for the compounds is 2 pmol. The method can be used for analysis of aspartylglycosylaminase activity in crude cell extracts and tissue samples.
Kaartinen
V
V
Department of Clinical Chemistry, Kuopio University Central Hospital, Finland.
Mononen
I
I
D000117:Acetylglucosamine / Q000031:analogs & derivatives / Q000302:isolation & purification / Q000378:metabolism; D000653:Amniotic Fluid / Q000201:enzymology; D001224:Aspartic Acid / Q000302:isolation & purification / Q000378:metabolism; D001227:Aspartylglucosylaminase / Q000097:blood / Q000378:metabolism*; D001921:Brain / Q000201:enzymology; D002233:Carbocysteine / Q000302:isolation & purification / Q000378:metabolism; D002823:Chorion / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005260:Female; D006801:Humans; D017879:Isothiocyanates; D007962:Leukocytes / Q000201:enzymology; D008168:Lung / Q000201:enzymology; D011247:Pregnancy; D012015:Reference Standards; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013861:Thiocyanates
D017879:Isothiocyanates; D013861:Thiocyanates; C005441:phenylisothiocyanate; C000009:N-acetylglucosaminylasparagine; D001224:Aspartic Acid; D002233:Carbocysteine; D001227:Aspartylglucosylaminase; D000117:Acetylglucosamine